DHA Induces Cell Death through the Production of ROS and the Upregulation of CHOP in Fibroblast-like Synovial Cells from Human Rheumatoid Arthritis Patients.

Rheumatoid arthritis (RA) is an inflammatory disease marked by a massive proliferation of synovial cells in the joints. In this study, we investigated the pro-apoptotic effects of docosahexaenoic acid (DHA) in human fibroblast-like synovial cells from RA patients (RA-FLS). An in vitro study using MH7A cells showed that DHA treatment induced caspase-8-dependent apoptosis in a dose-dependent manner and reduced the TNF-α-mediated induction of MMP-9 and IL-1ß. DHA also induced the phosphorylation of eIF2α, the expression of the ER stress markers ATF4 and C/EBP homologous protein (CHOP), and death receptor 5 (DR5). The knockdown of CHOP or DR5 increased cell viability and reduced apoptosis in DHA-treated cells. Furthermore, the knockdown of CHOP reduced DHA-mediated DR5 expression, while the overexpression of CHOP increased DR5 expression. We also found that DHA treatment induced the accumulation of reactive oxygen species (ROS), and pretreatment with the anti-oxidant Tiron effectively abrogated not only the expression of CHOP and DR5, but also DHA-induced apoptosis. Under this condition, cell viability was increased, while PARP-1 cleavage and caspase-8 activation were reduced. All the findings were reproduced in human primary synovial cells obtained from RA patients. These results suggest that the DHA-mediated induction of ROS and CHOP induced apoptosis through the upregulation of DR5 in RA-FLSs, and that CHOP could be used as a therapy for RA.

Dopa-responsive dystonia with a novel initiation codon mutation in the GCH1 gene misdiagnosed as cerebral palsy.

Dopa-responsive dystonia (DRD) is a clinical syndrome characterized by childhood-onset dystonia and a dramatic response to relatively low doses of levodopa. However, patients with DRD can be misdiagnosed as cerebral palsy or spastic diplegia due to phenotypic variation. Here we report a young woman with DRD who were severely disabled and misdiagnosed as cerebral palsy for over 10 yr. A small dose of levodopa restored wheelchair-bound state to normality. However, thoracolumbar scoliosis has remained as a sequel due to late detection of DRD. Genetic analysis by using PCR-direct sequencing revealed a novel initiation codon mutation (c.1A>T; p.Met1Leu) in GTP cyclohydrolase 1 (GCH1) gene. Although it is known that DRD can be misdiagnosed as cerebral palsy, this case reinforces the importance of differential diagnosis of DRD from cerebral palsy.

Salivary biomarkers in oral squamous cell carcinoma.

In disease diagnostics and health surveillance, the use of saliva has potential because its collection is convenient and noninvasive. Over the past two decades, the development of salivary utilization for the early detection of cancer, especially oral cavity and oropharynx cancer has gained the interest of the researcher and clinician. Until recently, the oral cavity and oropharynx cancers are still having a five-year survival rate of 62%, one of the lowest in all major human cancers. More than 90% of oral cancers are oral squamous cell carcinoma (OSCC). Despite the ease of accessing the oral cavity in clinical examination, most OSCC lesions are not diagnosed in the early stage, which is suggested to be the main cause of the low survival rate. Many studies have been performed and reported more than 100 potential saliva biomarkers for OSCC. However, there are still obstacles in figuring out the reliable OSCC salivary biomarkers and the clinical application of the early diagnosis protocol. The current review article discusses the emerging issues and is hoped to raise awareness of this topic in both researchers and clinicians. We also suggested the potential salivary biomarkers that are reliable, specific, and sensitive for the early detection of OSCC.

Preictal versus ictal injection of radiotracer for SPECT study in partial epilepsy: SISCOM.

A 27-year-old man had complex partial seizures and a dysembryoplastic neuroepithelial tumor (DNT) in the left inferior-basal temporal region. The patient's seizures consisted of incomprehensible speech, staring, unresponsiveness, fumbling and then looking around. For the brain SPECT study, radiotracer was injected during the preictal (11s prior to seizure onset), ictal (at 25 s out of 47 s seizure duration) and interictal periods. Interictal SPECT was subtracted from preictal or ictal-injection SPECTs and then the subtracted SPECTs were overlaid on the patient's MRI (SISCOM). SISCOM with preictal-injection SPECT showed hyperperfusion at the brain lesion, whereas SISCOM with ictal-injection SPECT showed hyperperfusion at the ipsilateral amygdala-hippocampus and hypoperfusion around the tumor lesion. After the DNT and nearby temporal lobe tissues were resected with preservation of amygdala-hippocampus, the patient became seizure free without complaint of subjective postsurgical memory decline. In this patient, SISCOM with preictal injection of radiotracer localized an epileptogenic zone, whereas SISCOM with the ictal injection showed hyperperfusion at the symptomatogenic zone.

Reduced cerebral blood flow during wakefulness in obstructive sleep apnea-hypopnea syndrome.

STUDY OBJECTIVES: To investigate changes in regional cerebral blood flow (rCBF) in patients with obstructive sleep apnea-hypopnea syndrome (OSAHS). DESIGN: We compared the 99mTc-ethylcysteinate dimer (ECD) single photon emission computed tomography (SPECT) images of patients with OSAHS with those of age- and sex-matched healthy volunteers. SETTING: University hospital. PATIENTS AND PARTICIPANTS: Twenty-seven patients with severe OSAHS and 27 healthy volunteers underwent 99mTc-ECD brain SPECT studies. INTERVENTION: For statistical parametric mapping analysis, all SPECT images were spatially normalized to the standard SPECT template and then smoothed using a 14-mm full-width at half-maximum Gaussian kernel. The Student t test was used for the statistical analysis. MEASUREMENTS AND RESULTS: The mean age of patients and subjects was 44.3 years (range 31-58). All patients underwent overnight polysomnography. The mean apnea-hypopnea index of patients was 60.4 +/- 17.6 per hour (range 33 -104), indicating severe OSAHS. All patients snored heavily and had daytime sleepiness (mean Epworth Sleepiness Scale score, 10.7 +/- 3.7, range 6-12). Statistical parametric mapping analysis showed that rCBF in patients with OSAHS was significantly reduced in bilateral parahippocampal gyri and in the right lingual gyrus, as compared with that of healthy volunteers (P < 0.05 with false discovery rate correction). Moreover, apnea-hypopnea indexes of patients were negatively correlated with rCBF in the right pericentral gyrus and right cuneus at uncorrected P < 0.001. CONCLUSIONS: Our results show the altered rCBF pattern in bilateral parahippocampal gyri, right lingual gyrus, pericentral gyrus, and cuneus in patients with severe OSAHS. These findings may partly explain the deficit in memory, spatial learning, executive function, and attention, which are frequently found in patients with OSAHS.

Antiepileptic effects of low-frequency repetitive transcranial magnetic stimulation by different stimulation durations and locations.

OBJECTIVE: To evaluate the antiepileptic effect of low-frequency rTMS (repetitive transcranial magnetic stimulation) in the patients with intractable epilepsy. METHODS: We enrolled 35 patients with localization-related epilepsy who had experienced at least one complex partial seizure or a secondarily generalized seizure per week on a constant antiepileptic drug regimen over an 8-week period. rTMS was administered using a Rapid(2) magnetic stimulator with an air-cooled coil at 0.5Hz for 5 consecutive days at 100% of rMT (resting motor threshold). Patients were divided into a focal stimulation group with a localized epileptic focus, or a non-focal stimulation group with a non-localized or multifocal epileptic focus. These two groups were then randomly subdivided into four subgroups depending on the total number of stimulations administered, i.e., 3000 pulse and 1500 pulse subgroups. Weekly seizure frequencies were determined for 8 weeks before and after rTMS. To compare the number of interictal spikes before and after rTMS, EEG was recorded twice before (1st day) and after rTMS (5th day). RESULTS: Mean weekly seizure frequency was non-significantly decreased after rTMS (8.4-->6.8/week, -13.9%). Longer stimulation subgroups (3000 pulses, -23.0%) tended to have fewer seizures than shorter stimulation subgroups (1500 pulses, -3.0%), without statistical significance. TMS stimulation site and structural brain lesions did not influence seizure outcome. However, interictal spikes significantly decreased (-54.9%, P=0.012) after rTMS and they totally disappeared in 6 patients (17.1%, 6/35). CONCLUSIONS: Low-frequency rTMS reduced interictal spikes, but its effect on seizure outcome was not significant. Focal stimulation for a longer duration tended to further reduce seizure frequency. SIGNIFICANCE: These findings may help clinicians to further investigate the therapeutic potential of the rTMS for patients with intractable epilepsy.

Neural correlates of musicogenic epilepsy: SISCOM and FDG-PET.

To localize the neural correlates of musicogenic epilepsy, subtraction ictal SPECT coregistered with MRI (SISCOM) and (18)F-fluorodeoxy glucose positron emission tomography (FDG-PET) were performed in a woman who had suffered from frequent musicogenic seizures. She had complex partial seizures consisting of palpitation and an unpleasant feeling, which were followed by staring and oroalimentary automatisms. Ictal EEG showed rhythmic theta waves originated from the right temporal lobe, and SISCOM showed ictal hyperperfusion on right insula, amygdala, hippocampal head, and anterior temporal lobe, whereas interictal FDG-PET showed interictal hypometabolism in the same brain regions, suggesting dysfunction and abnormal activation of right temporo-limbic structures related to an emotional response to music.

Structural brain abnormalities in juvenile myoclonic epilepsy patients: volumetry and voxel-based morphometry.

OBJECTIVE: We aimed to find structural brain abnormalities in juvenile myoclonic epilepsy (JME) patients. MATERIALS AND METHODS: The volumes of the cerebrum, hippocampus and frontal lobe and the area of the corpus callosum's subdivisions were all semiautomatically measured, and then optimized voxel-based morphometry (VBM) was performed in 19 JME patients and 19 age/gender matched normal controls. RESULTS: The rostrum and rostral body of the corpus callosum and the left hippocampus were significantly smaller than those of the normal controls, whereas the volume of the JME's left frontal lobe was significantly larger than that of the controls. The area of the rostral body had a significant positive correlation with the age of seizure onset (r = 0.56, p = 0.012), and the volume of the right frontal lobe had a significant negative correlation with the duration of disease (r = -0.51, p = 0.025). On the VBM, the gray matter concentration of the prefrontal lobe (bilateral gyri rectus, anterior orbital gyri, left anterior middle frontal gyrus and right anterior superior frontal gyrus) was decreased in the JME group (corrected p < 0.05). CONCLUSION: The JME patients showed complex structural abnormalities in the corpus callosum, frontal lobe and hippocampus, and also a decreased gray matter concentration of the prefrontal region, which all suggests there is an abnormal neural network in the JME brain.

Salubrinal-Mediated Upregulation of eIF2α Phosphorylation Increases Doxorubicin Sensitivity in MCF-7/ADR Cells.

Eukaryotic translation initiation factor 2 alpha (eIF2α), which is a component of the eukaryotic translation initiation complex, functions in cell death and survival under various stress conditions. In this study, we investigated the roles of eIF2α phosphorylation in cell death using the breast cancer cell lines MCF-7 and MCF-7/ADR. MCF-7/ADR cells are MCF-7-driven cells that have acquired resistance to doxorubicin (ADR). Treatment of doxorubicin reduced the viability and induced apoptosis in both cell lines, although susceptibility to the drug was very different. Treatment with doxorubicin induced phosphorylation of eIF2α in MCF-7 cells but not in MCF-7/ADR cells. Basal expression levels of Growth Arrest and DNA Damage 34 (GADD34), a regulator of eIF2α, were higher in MCF-7/ADR cells compared to MCF-7 cells. Indeed, treatment with salubrinal, an inhibitor of GADD34, resulted in the upregulation of eIF2α phosphorylation and enhanced doxorubicin-mediated apoptosis in MCF-7/ADR cells. However, MCF-7 cells did not show such synergic effects. These results suggest that dephosphorylation of eIF2α by GADD34 plays an important role in doxorubicin resistance in MCF-7/ADR cells.

Hempseed oil induces reactive oxygen species- and C/EBP homologous protein-mediated apoptosis in MH7A human rheumatoid arthritis fibroblast-like synovial cells.

ETHNOPHARMACOLOGICAL RELEVANCE: The medicinal efficacy of hempseed (Cannabis sativa L.), which is rich in polyunsaturated fatty acids, in atopic dermatitis, inflammation, and rheumatoid arthritis (RA) has been suggested for centuries. Hempseed has been used as a treatment for these diseases in Korean and Chinese folk medicine. The aim of the study is to investigate the effects of hempseed oil (HO) on MH7A human RA fibroblast-like synovial cells. MATERIALS AND METHODS: MH7A cells were used to study the anti-rheumatoid effects of hempseed (Cannabis sativa L., cv. Cheungsam/Cannabaceae) oil by investigating cell viability, apoptosis, lipid accumulation, oxidative stress, and endoplasmic reticulum (ER) stress-induced apoptosis. RESULTS: HO treatment reduced the survival rate of MH7A cells and promoted apoptotic cell death in a time- and dose-dependent manner. Both lipid accumulation and the level of intracellular reactive oxygen species (ROS) increased in HO-treated MH7A cells. Co-treatment with the antioxidant Tiron effectively abrogated the cytotoxic effects of HO; the ROS level was reduced, cell viability was recovered, and apoptotic cell death was significantly diminished. Moreover, HO-treated cells exhibited increased expression of the major ER stress markers, glucose-regulated protein 78 and C/EBP homologous protein (CHOP). The siRNA-mediated knockdown of CHOP prevented HO-induced apoptosis. CONCLUSIONS: Our results suggest that HO treatment induced lipid accumulation, ROS production, CHOP expression, and apoptosis in MH7A cells, and that CHOP functions as an anti-rheumatoid factor downstream of HO in MH7A cells.

Evaluation of brain iron content in idiopathic REM sleep behavior disorder using quantitative magnetic resonance imaging.

BACKGROUND: Neuroimaging studies in patients with idiopathic rapid eye movement sleep behavior disorder (iRBD) show similar structural and functional changes to alpha-synucleinopathies, including PD. Until now, there have been few attempts to characterize brain iron deposition in iRBD. The aim of this study was to investigate brain iron content in patients with iRBD using quantitative magnetic resonance imaging (MRI). METHODS: 3-T MRI was performed in 15 patients with iRBD and 20 age-matched healthy control subjects. In order to evaluate the iron-related neurodegenerative changes, we assessed volume and transverse relaxation rate (R2*) simultaneously. We used both region-based and voxel-based analysis. RESULTS: No significant differences in R2* values were found between iRBD groups and healthy control subjects. There were no areas of significantly reduced or increased gray matter and white matter volume in the iRBD group. Instead, lateral ventricle volumes measured automatically by FreeSurfer were significantly larger in patients with iRBD than in healthy controls (P < 0.05). CONCLUSION: The present study suggests that iron-related R2* values may not be an imaging biomarker for neurodegeneration in iRBD.

Obstructive hydrocephalus due to CNS toxocariasis.

A 46-year-old man developed intermittent headache, diplopia, and visual obscuration for two months. Funduscopic examination showed optic disk swelling in both eyes. Brain MRI exhibited hydrocephalus and leptomeningeal enhancement at the prepontine cistern, left cerebellopontine angle cistern and bilateral cerebral hemisphere, and hemosiderin deposition along the cerebellar folia. CSF analysis revealed an elevated opening pressure with xanthochromic appearance and small amount of red blood cells. Antibody titer against Toxocariasis using ELISA was elevated both in blood and CSF. Obstructive hydrocephalus and hemosiderin deposition in this case may result from the active inflammatory process due to CNS toxocariasis within the subarachnoid space.

Seizure duration determined by subdural electrode recordings in adult patients with intractable focal epilepsy.

BACKGROUND AND PURPOSE: To investigate the duration of seizures and its relationship to seizure type, epilepsy syndrome, and seizure clustering. METHODS: We examined 1,251 seizures from 152 patients who underwent video-electrocorticographic monitoring with subdural electrodes. Their seizure duration, seizure types, epilepsy syndromes, and seizure clusters were analyzed. RESULTS: The median seizure duration was 91.5s (4-1016s). There were 34 (2.7%) seizures lasting > 5 minutes in 20 (13.2%) patients. There was a significant difference in seizure duration according to seizure types (p < 0.0001), but not to epilepsy syndromes. There were 99 seizure clusters in 67 (44.1%) patients. The first seizure in a cluster of seizures tended to last longer than non-cluster seizures (median 98s versus 89s, p = 0.033). Seizure duration was significantly longer in mesial temporal lobe epilepsy than in neocortical lobe epilepsy (median 103s versus 87s, p = 0.041). Rate of seizure cluster was lower in mTLE (38.0%) than in NLE (47.1%), but this difference was not significant. CONCLUSIONS: Seizure durations were different among seizure types. Seizure clustering also differ between patients with mTLE and those with NLE, which suggests different seizure generation and propagation among different epileptogenic foci. This study has implications for the identification of abnormally prolonged seizures.

Cognitive effects of topiramate revealed by standardised low-resolution brain electromagnetic tomography (sLORETA) of event-related potentials.

OBJECTIVE: To evaluate the effect of topiramate (TPM) on event-related potentials (ERPs) in patients with epilepsy. METHODS: Neuropsychological tests and ERP study using auditory oddball paradigm were conducted before and after treatment with TPM in drug-naive epilepsy patients. To detect target brain regions in which ERP changed during the cognitive task, cortical current densities of ERP components were analysed using standardised low-resolution electromagnetic tomography (sLORETA). RESULTS: Neuropsychological tests (n=18 patients) showed that TPM significantly decreased the score in digit span, Corsi block and Controlled Oral Word Association word fluency. Repeated-measures analysis of variance of ERP data (n=13 patients) revealed that P2 amplitude was significantly increased at Fz electrode following treatment with TPM. Statistical non-parametric map of sLORETA between pre- and post-TPM ERPs revealed that current density of P200 component was significantly reduced by TPM in bilateral parieto-occipital, temporolimbic and dorsolateral right prefrontal regions. CONCLUSIONS: Our findings suggest that TPM affects selective brain regions which may be related to cognitive side effects. SIGNIFICANCE: Source localisation of ERPs can be helpful in identifying target brain regions for the cognitive side effects of anti-epileptic drugs.

Contralateral hyperperfusion and ipsilateral hypoperfusion by ictal SPECT in patients with mesial temporal lobe epilepsy.

Ictal Single Proton Emission Computed Tomography (SPECT) has demonstrated high levels of sensitivity in localizing seizures among patients with epilepsy of the mesial temporal lobe (mTLE). However, incorrect information on the lateralization of mTLE has also been reported. In order to investigate the causes of these incorrect localizations, the authors assessed clinical symptoms, as well as the electroencephalography (EEG) and brain SPECT scan data of five patients with mTLE experiencing ictal hyperperfusion of the contralateral temporal lobe. All patients underwent presurgical evaluations, including an interictal and ictal brain SPECT scan. A subtraction ictal SPECT co-registered with Magnetic Resonance Imaging (MRI) procedure or SISCOM was performed. Hyperperfusion (ictal perfusion greater than interictal perfusion) and hypoperfusion (ictal perfusion lower than interictal perfusion), results of SISCOM were analyzed and compared with seizure and ictal EEG pattern patterns. All the five patients had unilateral hippocampal sclerosis, and the radiotracer for the ictal SPECT was injected after the ictal EEG pattern had propagated to the contralateral side. The average delay between the ictal EEG onset and the radiotracer injection was 29.7+/-9.6s. All hyperperfusion SISCOM results revealed hyperperfusion in the contralateral temporal region with a more intense ictal EEG build-up. However, hypoperfusion SISCOM results demonstrated significant hypoperfusion in the epileptogenic temporal lobe of three of the five patients, but no hypoperfusion finding in the other two patients. This study demonstrates that early ictal EEG pattern propagation to the contralateral side in mTLE may be associated with contralateral ictal hyperperfusion with or without ipsilateral temporal hypoperfusion. The authors recommend simultaneous interpretations of ictal SPECT and ictal EEG propagation patterns at the time of the injection of radiotracers.

Disabling Head Tremor in a Patient with DYT1 Mutation.

Dystonic head tremor is known to be a feature in some patients with DYT1 mutation. However, isolated tremor of the head without relevant cervical dystonia has rarely been described. We report here a patient with the three-bp GAG deletion in the DYT1 gene (904_906delGAG) who had severe head tremor in the frame of a generalized limb dystonia.

Apigenin-induced apoptosis is mediated by reactive oxygen species and activation of ERK1/2 in rheumatoid fibroblast-like synoviocytes.

Fibroblast-like synovial cells play a crucial role in the pathophysiology of rheumatoid arthritis (RA), as these cells are involved in inflammation and joint destruction. Apigenin, a dietary plant-flavonoid, is known to have many functions in animal cells including anti-proliferative and anticancer activities, but its role in human rheumatoid arthritis fibroblast-like synoviocytes (RA-FLSs) has not been reported. In this study, we investigated the roles of apigenin in RA-FLSs. The survival rate decreased, and apoptotic cell death was induced by apigenin treatment in RA-FLSs. Apigenin treatment resulted in activation of the mitogen-activated protein kinase (MAPK) ERK1/2, and pretreatment with an ERK inhibitor PD98059 dramatically reduced apigenin-induced apoptosis. We found that apigenin-mediated production of a large amount of intracellular reactive oxygen species (ROS) caused activation of ERK1/2 and apoptosis; treatment with the antioxidant Tiron strongly inhibited the apigenin-induced generation of ROS, phosphorylation of ERK1/2, and apoptotic cell death. Apigenin-induced apoptotic cell death was mediated through activation of the effectors caspase-3 and caspase-7, and was blocked by pretreatment with Z-VAD-FMK (a pan-caspase inhibitor). These results showed that apigenin-induced ROS and oxidative stress-activated ERK1/2 caused apoptotic cell death in apigenin-treated RA-FLSs.

Can pentobarbital replace amobarbital in the Wada test?

To investigate the usefulness of pentobarbital (PTB) in the Wada test, 32 patients injected with PTB and 28 patients injected with amobarbital (AMB) were retrospectively analyzed. The AMB and PTB groups did not significantly differ with respect to mean time for recovery to grade III or V motor activity and duration of EEG delta slowing. The incidence of drowsiness or confusion after injection was lower in the PTB group (P=0.043). Language lateralization was well established in both groups. Fifty-three percent of patients in the PTB group and 46.2% in the AMB group with unilateral temporal lobe epilepsy had memory dominance in the nonepileptic hemisphere. The usefulness of PTB in terms of language and memory lateralization was found to be equivalent to that of AMB. Moreover, PTB produced drowsiness and confusion less frequently than AMB, although one patient in the PTB group experienced transient respiratory depression without any sequelae.